A case of ocular infection with Onchocerca lupi in a dog from Germany.

Onchocerca lupi is an emerging zoonotic parasite infecting the ocular connective tissue of dogs, cats and humans. The only known case of canine ocular onchocerciasis in Germany was documented in 2002 in a shelter dog. However, the species of Onchocerca causing the infection could not be identified. Here, we report a case of the ocular infection with O. lupi in a dog, confirmed by PCR and sequencing of the cox1 gene. Further investigations are required to assess the risk factors for transmission and spread of the parasite in Germany.

Diagnostic exome sequencing of syndromic epilepsy patients in clinical practice.

Although genetic revolution of recent years has vastly expanded a list of genes implicated in epilepsies, complex architecture of epilepsy genetics is still largely unknown, consequently, universally accepted workflows for epilepsy genetic testing in a clinical practice are missing. We present a comprehensive NGS-based diagnostic approach addressing both the clinical and genetic heterogeneity of disorders involving epilepsy or seizures. A bioinformatic panel of 862 epilepsy- or seizure-associated genes was applied to Mendeliome (4813 genes) or whole-exome sequencing data as a first stage, while the second stage included untargeted variant interpretation. Eighty-six consecutive patients with epilepsy or seizures associated with neurodevelopmental disorders and/or congenital malformations were investigated. Of the 86 probands, 42 harbored pathogenic and likely pathogenic variants, giving a diagnostic yield of 49%. Two patients were diagnosed with pathogenic copy number variations and 2 had causative mitochondrial DNA variants. Eleven patients (13%) were diagnosed with diseases with specific treatments. Besides, genomic approach in diagnostics had multiple additional benefits due to mostly non-specific, overlapping, not full-blown phenotypes and abilities to diagnose novel and ultra rare epilepsy-associated diseases. Likely pathogenic variants were identified in SOX5 gene, not previously associated with epilepsy, and UBA5, a recently associated with epilepsy gene.

Genetic variation in leptin and leptin receptor genes as a risk factor for idiopathic male infertility.

The aim of this study was to examine whether there is an association among genetic variability in leptin (LEP) and leptin receptor (LEPR) genes and male infertility. We performed a case-control study and were searching for an association between polymorphisms of LEP and LEPR genes and male infertility. The study group consisted of 317 patients with idiopathic infertility and a control group of 241 fertile men from Slovenia. Four single nucleotide polymorphisms (SNPs) in LEP gene and four single nucleotide polymorphisms (SNPs) in LEPR gene were chosen and genotyped. Statistically significant SNP was further validated in additional 255 infertile patients and 168 controls from Serbia and Macedonia. In the Slovenian population, we found a statistically significant difference in genotype distribution for rs10244329 polymorphism in LEP gene (recessive genotype model, p value = 0.048). The trend toward statistically significant difference in genotype distribution for rs10244329 polymorphism was confirmed in the Serbian and Macedonian populations (p value = 0.07). Our data suggest that genetic variability in the LEP gene might be associated with male infertility warranting further confirmation and mechanistic investigations.

Occurrence of ticks in the subcutaneous tissue of red foxes, Vulpes vulpes in Czech Republic and Romania.

An extensive survey of parasites in red foxes (Vulpes vulpes) was independently conducted in Romania and the Czech Republic. Carcasses were examined by necropsy, and small, dark nodules apparently containing ticks were noticed in the subcutaneous tissue of several foxes. Histopathological examination was performed using hematoxylin and eosin (HE) staining. Of the 91 foxes examined from the Czech Republic, 14 (15.4%) were harboring ticks in the subcutaneous tissue. In the majority of these cases, 1-3 nodules/fox were found, with a maximum of 31 nodules/fox. In Romania a single examined fox had subcutaneous ticks. All ticks collected from subcutaneous tissue were partially engorged adults. Based on morphological features, Ixodes ricinus, I. hexagonus, I. crenulatus and Dermacentor reticulatus were identified. The histopathological examination revealed chronic granulomatous panniculitis with peripheral fibrosis and intralesional presence of the ticks. Only few data are available regarding ticks localized in the subcutaneous tissue of any host. All the ticks were dead or already decomposed and it is evident that subcutaneous location does not represent an evolutionary advantage, as the detachment and finishing the life cycle is impossible.

First Molecular Evidence of Zoonotic Bacteria in Ticks in Bosnia and Herzegovina.

In Bosnia and Herzegovina, the tick fauna is very diverse, but data on the occurrence of zoonotic tick-borne bacteria are lacking. Thus, the aim of this study was to investigate the presence of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, 'Candidatus Neoehrlichia mikurensis', spotted fever group (SFG) rickettsiae and Francisella tularensis in questing ticks. In 19 (21.8%) of 87 ticks (Ixodes ricinus, n = 30; Dermacentor reticulatus, n = 54; D. marginatus, n = 3) collected by flagging the vegetation at the collection site in the Glamoc Municipality (south-western Bosnia and Herzegovina), Rickettsia monacensis (1.1%), R. helvetica (5.7%), R. raoultii (5.7%), R. slovaca (8.0%), A. phagocytophilum (1.1%) and F. tularensis subsp. holartica (1.1%) were detected and identified by molecular methods. None of the tested ticks were positive for B. burgdorferi s.l. and 'Candidatus N. mikurensis', and co-infection of R. slovaca and F. tularensis subsp. holarctica was detected in only one D. marginatus (1.1%). This study reports the occurrence of emerging zoonotic bacteria in ticks from Bosnia and Herzegovina for the first time, indicating a public health threat to humans. Therefore, physicians and practitioners should be aware of the presence of these tick-borne bacteria, especially when they are faced with acute febrile illnesses after tick exposure.

Non-linear partitioning and organic volatility distributions of urban aerosols.

Gas-to-particle partitioning of organic aerosols (OA) is represented in most models by Raoult's law, and depends on the existing mass of particles into which organic gases can dissolve. This raises the possibility of non-linear response of particle-phase OA mass to the emissions of precursor volatile organic compounds (VOCs) that contribute to this partitioning mass. Implications for air quality management are evident: a strong non-linear dependence would suggest that reductions in VOC emission would have a more-than-proportionate benefit in lowering ambient OA concentrations. Chamber measurements on simple VOC mixtures generally confirm the non-linear scaling between OA and VOCs, usually stated as a mass-dependence of the measured OA yields. However, for realistic ambient conditions including urban settings, no single component dominates the composition of the organic particles, and deviations from linearity are presumed to be small. Here we re-examine the linearity question using volatility spectra from several sources: (1) chamber studies of selected aerosols, (2) volatility inferred for aerosols sampled in two megacities, Mexico City and Paris, and (3) an explicit chemistry model (GECKO-A). These few available volatility distributions suggest that urban OA may be only slightly super-linear, with most values of the normalized sensitivity exponent in the range 1.1-1.3, also substantially lower than seen in chambers for some specific aerosols. The rather low exponents suggest that OA concentrations in megacities are not an inevitable convergence of non-linear effects, but can be addressed (much like in smaller urban areas) by proportionate reductions in emissions.

Influence of Fasciola hepatica on Serum Biochemical Parameters and Vascular and Biliary System of Sheep Liver.

BACKGROUND: The aim of this study was to evaluate the functional capacity of the liver based on the activity of specific enzymes and bilirubin in serum and also to investigate the influence of mechanical and toxic effects of Fasciola hepatica on the structures of the blood vessels and biliary tract in the sheep liver. METHODS: Blood samples and liver of 63 indigenous sheep of Pramenka breed, slaughtered in the period from March to December 2009 were used. Based on parasitological findings in the liver, all animals were divided into two groups: control (n = 34) and infected group (n = 29). For investigation and description of pathological changes in sheep liver, naturally infected with F. hepatica, corrosion cast technique was used. RESULTS: Biochemical analysis of tested parameters showed a significant elevation (P≤0.05) of serum gamma-glutamyl transferase (GGT), total bilirubin (TBIL) and direct bilirubin (DBIL) in infected sheep group comparing with the control group. No significant differences were observed for activity of aspartate aminotranferase (AST) between groups. Vascular and biliary systems of the liver were found to be affected. CONCLUSION: Results of biochemical analysis are consistent with pathological findings and measuring of tested parameters could be used in early diagnosis of sheep fasciolosis and to test the effectiveness of anthelmintic therapy. Corrosion cast technique is very useful for investigation of pathological changes and neoangiogenesis of vascular and biliary system in sheep liver, caused by mechanical and toxic effects of F. hepatica.

Fatal verminous pharyngitis and esophagitis caused by Streptocara incognita in mute swans (Cygnus olor).

Streptocara spp. infections are reported to cause gastritis, proventriculitis, esophagitis, and pharyngitis in various waterfowls, especially diving ducks. In the present paper, we describe severe fatal diphtheritic pharyngitis and esophagitis caused by Streptocara incognita in three female mute swans (Cygnus olor) in Bosnia and Herzegovina. Prior to death, the swans were showing signs of lethargy, anorexia, and reluctance to move. At necropsy, in all swans severe diphtheritic pharyngitis and esophagitis with deep, dark red hemorrhagic ulcerations were observed. Numerous thin, white, up to 1-cm-long nematodes, identified as S. incognita, were observed embedded in the pharyngeal and esophageal mucosa under the diphtheritic membranes. Histopathology revealed severe fibrinonecrotic inflammation with numerous cross-sections of the parasites. To the authors' knowledge, this is the first report of severe, fatal streptocariasis in mute swans.

Three dimensional echocardiographic assessment of multiple rhabdomyoma in newborn.

BACKGROUND: Cardiac rhabdomyomas represent the most common primary cardiac tumour in children and are strongly associated with tuberous sclerosis complex. RESULTS: We reported a newborn for whom three-dimensional echocardiogram, with multiplane mode, real-time imaging, full volume and i-slice view, allowed detailed visualisation of multiple highly echogenic and well-circumscribed cardiac rhabdomyoma. CONCLUSIONS: Three-dimensional imaging allowed a better definition of the tumour characteristics and provided a better delineation of the spatial relationship of the mass with a tomographic perspective. Three dimensional imaging may facilitate a possible operative planning and should be included in cardiac mass evaluation and follow-up.

Losartan's affinity to fluid bilayers modulates lipid-cholesterol interactions.

Losartan is an angiotensin II receptor antagonist mainly used for the regulation of high blood pressure. Since it was anticipated that losartan reaches the receptor site via membrane diffusion, the impact of losartan on model membranes has been investigated by small angle X-ray scattering. For this purpose 2-20 mol% losartan was incorporated into dimyristoyl-phosphatidylcholine (DMPC) and palmitoyl-oleoyl-phosphatidylcholine (POPC) bilayers and into their binary mixtures with cholesterol in the concentration range of 0 to 40 mol%. Effects of losartan on single component bilayers are alike. Partitioning of losartan into the membranes confers a negative charge to the lipid bilayers that causes the formation of unilamellar vesicles and a reduction of the bilayer thickness by 3-4%. Analysis of the structural data resulted in an estimate for the partial area of losartan, A(Los) ≈ 40 Å(2). In the presence of cholesterol, differences between the effects of losartan on POPC and DMPC are striking. Membrane condensation by cholesterol is retarded by losartan in POPC. This contrasts with DMPC, where an increase of the cholesterol content shifts the partitioning equilibrium of losartan towards the aqueous phase, such that losartan gets depleted from the bilayers from 20 mol% cholesterol onwards. This indicates (i) a chain-saturation dependent competition of losartan with lipid-cholesterol interactions, and (ii) the insolubility of losartan in the liquid ordered phase of PCs. Consequently, losartan's action is more likely to take place in fluid plasma membrane patches rather than in domains rich in cholesterol and saturated lipid species such as in membrane rafts.

Impact of trash burning on air quality in Mexico City.

Air pollution experienced by expanding urban areas is responsible for serious health effects and death for millions of people every year. Trash burning is a common disposal method in poor areas, yet it is uncontrolled in many countries, and its contribution to air pollution is unclear due to uncertainties in its emissions. Here we develop a new trash burning emission inventory for Mexico City based on inverse socioeconomic levels and recently measured emission factors, and apply a chemistry-transport model to analyze the effects on pollutant concentrations. Trash burning is estimated to emit 25 tons of primary organic aerosols (POA) per day, which is comparable to fossil fuel POA emissions in Mexico City, and causes an increase in average organic aerosol concentrations of ∼0.3 µg m(-3) downtown and up to 2 µg m(-3) in highly populated suburbs near the sources of emission. An evaluation using submicrometer antimony suggests that our emission estimates are reasonable. Mitigation of trash burning could reduce the levels of organic aerosols by 2-40% and those of PM(2.5) by 1-15% over the metropolitan area. The trash burning contributions to carbon monoxide, nitrogen oxides, and volatile organic compounds were found to be very small (<3%), and consequently the contributions to secondary nitrate, sulfate, and secondary organic aerosols are also very small.

Small- and wide-angle X-ray scattering (SWAXS) for quantification of aspirin content in a binary powder mixture.

This article presents a novel application of small and wide angle X-ray scattering (SWAXS) in the assessment of aspirin and lactose content in a binary pharmaceutical powder formulation. It is shown that the content correlates with the intensity of the SAXS signal and the intensity of polymorph fingerprints in the WAXS spectra that are collected from the same samples. Because the polymorph WAXS fingerprints and the SAXS signal are two independent characteristics of the same sample, simultaneous SWAXS analysis provides the basis for a dual independent assessment of the same contents.

Interactions of the AT1 antagonist valsartan with dipalmitoyl-phosphatidylcholine bilayers.

Valsartan is a marketed drug with high affinity to the type 1 angiotensin (AT1) receptor. It has been reported that AT1 antagonists may reach the receptor site by diffusion through the plasma membrane. For this reason we have applied a combination of differential scanning calorimetry (DSC), Raman spectroscopy and small and wide angle X-ray scattering (SAXS and WAXS) to investigate the interactions of valsartan with the model membrane of dipalmitoyl-phosphatidylcholine (DPPC). Hence, the thermal, dynamic and structural effects in bulk as well as local dynamic properties in the bilayers were studied with different valsartan concentrations ranging from 0 to 20 mol%. The DSC experimental results showed that valsartan causes a lowering and broadening of the phase transition. A splitting of the main transition is observed at high drug concentrations. In addition, valsartan causes an increase in enthalpy change of the main transition, which can be related to the induction of interdigitation of the lipid bilayers in the gel phase. Raman spectroscopy revealed distinct interactions between valsartan with the lipid interface localizing it in the polar head group region and in the upper part of the hydrophobic core. This localization of the drug molecule in the lipid bilayers supports the interdigitation view. SAXS measurements confirm a monotonous bilayer thinning in the fluid phase, associated with a steady increase of the root mean square fluctuation of the bilayers as the valsartan concentration is increased. At high drug concentrations these fluctuations are mainly governed by the electrostatic repulsion of neighboring membranes. Finally, valsartans' complex thermal and structural effects on DPPC bilayers are illustrated and discussed on a molecular level.

The influence of dietary palm olein, fish oil and lard on the egg yolk and plasma lipid composition, and performances of laying hens.

The influence of dietary palm olein, in comparison to fish oil and lard, on lipid levels in egg yolk and blood plasma, the fatty acid composition of egg yolk, and various production parameters were studied. Brown Lohman laying hens (n=45) were randomly assigned into three groups of 15 birds, and treated with experimental diets with either 3% palm olein (PO), fish oil (FO) or lard (L) for 6 weeks. At the end of the experiment, feed consumption was significantly lower for hens fed the PO diet, except week 6 of the experiment. The concentration of plasma triglycerides was increased by all experimental diets, whereas there were no significant increases of plasma total lipid and total cholesterol concentrations only in the PO group. For yolk lipids a decrease in triglycerides in the FO and L groups was observed, while total cholesterol and total lipid were significantly decreased in the PO group. Feeding with the PO diet resulted in the lowest concentrations of palmitic, stearic and linoleic acid, as well as in the highest concentration of monounsaturated oleic acid in the yolk total lipid. It was concluded that the composition of yolk lipids did not closely match the concentrations of lipids observed in experimental diets or plasma. Based on the current work it seems that the PO diet modulates egg yolk lipid content best.

Genetic diversity and structure of the West Balkan Pramenka sheep types as revealed by microsatellite and mitochondrial DNA analysis.

Several different phenotypes of the native Pramenka sheep have been developed in the Balkan region for different environmental and socio-cultural conditions. Animals from seven West Balkan Pramenka sheep types were analysed for 15 microsatellite markers and for mitochondrial DNA (mtDNA) and the results were used to assess genetic variation within and among the types and to infer the genetic population structure of the Pramenka sheep. Mean expected heterozygosity and allelic richness over the microsatellite loci and sheep types were 0.78 and 7.9, respectively. A Bayesian statistical method for estimating hidden genetic structure suggested that a core of the largest panmictic population was formed by Serbian, Kosovan, Bosnian, Montenegrin and Albanian types, while Croatian and Macedonian types comprised two other main populations, respectively. Mitochondrial DNA analysis revealed two mtDNA haplogroups in the Pramenka sheep, B and A, with a frequency of 93.7% and 6.3%, respectively. A total of 60 mtDNA haplotypes were found in 64 animals sequenced, and the mean nucleotide and haplotypic diversities over the types were 0.013 and 0.945, respectively. Molecular analysis suggests that the West Balkan Pramenka sheep types have their origins in two distinct maternal lineages of domestic sheep and different Pramenka phenotypes tend to form few panmictic populations. The Pramenka sheep represents a valuable resource of genetic diversity in sheep.